Fig. 6.

Compound C inhibits adipogenesis of C3H10T1/2 cells.

A) Confluent CH3H10T1/2 cells were stimulated for 5 days with IID, in the presence or absence of the AMPK inhibitor, 10 μM compound C. Cell extracts were then prepared and immunoblotted with antibodies to phosphorylated ACC (Ser 79). Representative immunoblots from an experiment carried out on three separate occasions with similar results are shown (upper panel). Densitometric values from 3 separate experiments are shown in the lower panel as means ± SEM. Significant decreases relative to control are indicated, #, p < 0.05.

B) Confluent CH3H10T1/2 cells were treated with 10% FCS supplemented with adipogenic cocktail (IID), in the presence or absence of the indicated concentrations of compound C. After 5 days cells were fixed with formalin and stained with Oil Red O to detect neutral lipid accumulation. Representative micrographs from an experiment carried out on three separate occasions with similar results are shown.

C) Confluent CH3H10T1/2 MSCs were induced to differentiate by addition 10% FCS in the presence or absence of IID medium and/or 10 μM Compound C or 10 μM rapamycin. Cell extracts were then prepared after 5 days and immunoblotted with antibodies to perilipin and tubulin. Representative immunoblots from an experiment carried out on three separate occasions with similar results are shown. Densitometric analysis of three immunoblots are shown as means ± SEM in the lower panel. Significant increases relative to control are indicated, ***p < 0.001 and significant decreases with respect to IID-treated cells are indicated, ###, p < 0.001.